Transforming growth factor-β1 induces Nox4 NAD(P)H oxidase and reactive oxygen species-dependent proliferation in human pulmonary artery smooth muscle cells
Abstract
Transforming growth factor-β1 (TGF-β1) is abundantly expressed in pulmonary hypertension, but its effect on the pulmonary circulation remains unsettled. We studied the consequences of TGF-β1 stimulation on freshly isolated human pulmonary artery smooth muscle cells (HPASMC). TGF-β1 initially promoted differentiation, with upregulated expression of smooth muscle contractile proteins. TGF-β1 also induced expression of Nox4, the only NAD(P)H oxidase membrane homolog found in HPASMC, through a signaling pathway involving Smad 2/3 but not mitogen-activated protein (MAP) kinases. TGF-β1 likewise increased production of reactive oxygen species (ROS), an effect significantly reduced by the NAD(P)H oxidase flavoprotein inhibitor diphenylene iodonium (DPI) and by Nox4 siRNAs. In the absence of TGF-β1, Nox4 was present in freshly cultured cells but progressively lost with each passage in culture, paralleling a decrease in ROS production by HPASMC over time. At a later time point (72 h), TGF-β1 promoted HPASMC proliferation in a manner partially inhibited by Nox4 small interfering RNA and dominant negative Smad 2/3, indicating that TGF-β1 stimulates HPASMC growth in part by a redox-dependent mechanism mediated through induction of Nox4. HPASMC activation of the MAP kinases ERK1/2 was reduced by the NAD(P)H oxidase inhibitors DPI and 4-(2-aminoethyl)benzenesulfonyl fluoride, suggesting that TGF-β1 may facilitate proliferation by upregulating Nox4 and ROS production, with transient oxidative inactivation of phosphatases and augmentation of growth signaling cascades. These findings suggest that Nox4 is the relevant Nox homolog in HPASMC. This is the first observation that TGF-β1 regulates Nox4, with important implications for mechanisms of pulmonary vascular remodeling.
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NOS are dimeric, multidomain enzymes that synthesize NO from molecular oxygen and l-arginine and use heme, tetrahydrobiopterin, FAD, FMN, and NADPH as cofactors.
“NOS are dimeric, multidomain enzymes that synthesize NO from molecular oxygen and l-arginine and use iron protoporphyrin-IX (heme), tetrahydrobiopterin (BH4), FAD, flavin mononucleotide (FMN), and NADPH as cofactors”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [904]
The canonical pathways of NO formation rely on the specialized NOS enzymes.
“The canonical pathways of NO formation rely on the specialized NOS enzymes.”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [904]
BH4 plays a critical role in the catalytic cycle of NOS.
“In addition, BH4 plays a critical role in the catalytic cycle of NOS (87, 204, 377, 904, 933, 1000, 1007).”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [904]
In saturating conditions of the NOS cofactors, the synthesis of NO only requires l-arginine and O2 as substrates.
“In saturating conditions of the NOS cofactors, the synthesis of NO only requires l-Arginine and O2 as substrates (904).”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [904]
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- Authors
- Sturrock, A., Cahill, B., Norman, K., Huecksteadt, T. P., Hill, K., Sanders, K., Karwande, S. V., Stringham, J. C., Bull, D. A., Gleich, M., Kennedy, T. P., & Hoidal, J. R.
- Affiliations
- University of Utah (US)
- Venue
- American Journal of Physiology-Lung Cellular and Molecular Physiology
- Publisher
- American Physiological Society
- Published
- 2006-04
- Volume
- 290
- Issue
- 4
- Pages
- L661-L673
- ISSN
- 1040-0605 (ISSN-L)
- 1522-1504
- OpenAlex
- W2131484844
- Cited by
- 402
- References
- 44
- Citations by year
- 2026: 6 · 2025: 5 · 2024: 9 · 2023: 10 · 2022: 17 · 2021: 10 · 2020: 13 · 2019: 17 · 2018: 21 · 2017: 29 · 2016: 23 · 2015: 29 · 2014: 33 · 2013: 28 · 2012: 29
- FWCI
- 14.44
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- American Journal of Physiology-Lung Cellular and Molecular Physiology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Heart, Lung, and Blood Institute (R01HL067281, HL67281)
- Topics
- Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Pulmonary Hypertension Research and Treatments
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › TGF-β signaling in diseases
- Health Sciences › Medicine › Physiology › Nitric Oxide and Endothelin Effects
- Keywords
- TGF-β1, NOX4, NADPH oxidase, reactive oxygen species, pulmonary artery smooth muscle cells, cell proliferation, Smad2/3 signaling, pulmonary hypertension, pulmonary vascular remodeling, ERK1/2 MAP kinase, diphenyleneiodonium, smooth muscle differentiation
- MeSH terms
- NADPH Oxidase 4, Cells, Cultured, Contractile Proteins, Humans, Pulmonary Artery, Signal Transduction, Transforming Growth Factor beta, Reactive Oxygen Species, NADPH Oxidases, Mitogen-Activated Protein Kinases, Myocytes, Smooth Muscle, Cell Proliferation, Smad Proteins, Transforming Growth Factor beta1
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